EP3919488B1 - Immunmodulator - Google Patents

Immunmodulator

Info

Publication number
EP3919488B1
EP3919488B1 EP20748337.1A EP20748337A EP3919488B1 EP 3919488 B1 EP3919488 B1 EP 3919488B1 EP 20748337 A EP20748337 A EP 20748337A EP 3919488 B1 EP3919488 B1 EP 3919488B1
Authority
EP
European Patent Office
Prior art keywords
mmol
added
ethyl
methyl
compound
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
EP20748337.1A
Other languages
English (en)
French (fr)
Other versions
EP3919488C0 (de
EP3919488A1 (de
EP3919488A4 (de
Inventor
Jin Li
Dengyou ZHANG
Fei Pan
Rong MA
Wenji ZHU
Yanfei XIN
Si Li
Weiping Liu
Yanping LIN
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hitgen Inc
Original Assignee
Hitgen Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hitgen Inc filed Critical Hitgen Inc
Publication of EP3919488A1 publication Critical patent/EP3919488A1/de
Publication of EP3919488A4 publication Critical patent/EP3919488A4/de
Application granted granted Critical
Publication of EP3919488C0 publication Critical patent/EP3919488C0/de
Publication of EP3919488B1 publication Critical patent/EP3919488B1/de
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4188—1,3-Diazoles condensed with other heterocyclic ring systems, e.g. biotin, sorbinil
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5386—1,4-Oxazines, e.g. morpholine spiro-condensed or forming part of bridged ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/08—Bridged systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • C07D491/107—Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/08—Bridged systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants

Definitions

  • the invention relates to an immunomodulator, in particular to a class of compounds that activate STING and its use as an immunomodulator in the manufacture of a medicament.
  • the human body's immune system can generally be divided into “natural immunity” system and “adaptive immunity” system.
  • the natural immune system plays an important role in resisting infections, inhibiting tumor growth and the pathogenesis of autoimmune diseases. It mainly recognizes pathogenic microorganisms and cancer cell components through pattern recognition receptors, initiates downstream signal pathways, and finally induces cytokine expression, kill pathogenic microorganisms and cancer cell components, and adapt to the immune system to promote the production of antibodies and specific T lymphocytes.
  • STING interferon gene stimulating factor, TMEM173, MITA, etc.
  • TMEM173, MITA interferon gene stimulating factor
  • Compounds that induce human interferon can be used to treat various diseases (including allergic diseases and other inflammatory diseases, allergic rhinitis and asthma, infectious diseases, neurodegenerative diseases, precancerous syndrome and cancer), and can also be used as immunization composition or vaccine adjuvant. Therefore, the development of new compounds that are capable of activating STING is an effective method for the treatment of type 1 IFN pathway diseases (including inflammatory diseases, allergic and autoimmune diseases, infectious diseases, cancer, and precancerous syndromes).
  • WO2017175147 discloses heterocyclic amides that are useful as modulators of STING.
  • the present invention provides a compound, or the stereoisomer, or the pharmaceutically acceptable salt thereof, wherein the compound is represented by formula II as defined by claim 1
  • the present invention further provides, a compound, or the stereoisomer, or the pharmaceutically acceptable salt thereof, wherein the compound is:
  • the present invention further provides a compound, or the stereoisomer, or the pharmaceutically acceptable salt thereof, wherein the compound is represented by formula III as defined in claim 3.
  • the present invention further provides the compound, or the stereoisomer, or the pharmaceutically acceptable salt thereof, wherein the compound is:
  • the present invention also provides the above-mentioned compound, or a stereoisomer, or a pharmaceutically acceptable salt thereof for use in treating diseases related to STING activity that are one or more of diseases related to inflammatory diseases, autoimmune diseases, infectious diseases, cancer, and precancerous syndrome
  • the present invention also provides the above-mentioned compound, or a stereoisomer, or a pharmaceutically acceptable salt thereof for use as an immune adjuvant.
  • the present invention also provides a medicament, which is a preparation prepared from the above-mentioned compound, or a stereoisomer, or a pharmaceutically acceptable salt thereof as the active ingredient, plus pharmaceutically acceptable auxiliary materials.
  • the diseases related to STING activity defined in the present invention are diseases in which STING plays an important role in the pathogenesis of the disease.
  • Diseases related to STING activity include inflammatory diseases, allergic diseases and autoimmune diseases, infectious diseases, cancer, and precancerous syndrome.
  • Cancer or “malignant tumor” refers to any of a variety of diseases characterized by uncontrolled abnormal cell proliferation, and the ability of affected cells to spread to other locations locally or through the bloodstream and lymphatic system of the body (i.e., metastasis) and any of many characteristic structural and/or molecular characteristics.
  • Cancer cells refer to cells that undergo multiple stages of tumor progression in the early, middle, or late stages. Cancers include sarcoma, breast cancer, lung cancer, brain cancer, bone cancer, liver cancer, kidney cancer, colon cancer, and prostate cancer.
  • the compound of formula I is used to treat a cancer selected from colon cancer, brain cancer, breast cancer, fibrosarcoma, and squamous cell carcinoma.
  • the cancer is selected from melanoma, breast cancer, colon cancer, lung cancer, and ovarian cancer.
  • the cancer being treated is a metastatic cancer.
  • Inflammatory diseases include a variety of conditions characterized by histopathological inflammation.
  • inflammatory diseases include acne vulgaris, asthma, enterocoelia diseases, chronic prostatitis, glomerulonephritis, inflammatory bowel disease, pelvic inflammation, reperfusion injury, rheumatoid arthritis, sarcoidosis, vasculitis, airway inflammation and interstitial cystitis caused by house dust mites.
  • Some embodiments of the present invention relate to the treatment of asthma, an inflammatory disease.
  • the immune system is usually involved in inflammatory diseases, which are manifested in allergic reactions and some myopathy. Many immune system diseases lead to abnormal inflammation.
  • radical groups or terms are applicable to the radical groups or terms throughout the specification; for terms not specifically defined herein, it should be, based on the disclosure and context, given the meaning that those skilled in the art can give them.
  • substitution refers to the replacement of a hydrogen atom in a molecule by a different atom or molecule.
  • the minimum and maximum content of carbon atoms in a hydrocarbon group is indicated by a prefix.
  • the prefix (C a -C b ) alkyl indicates any alkyl group containing "a" to "b” carbon atoms.
  • (C 1 -C 6 ) alkyl refers to an alkyl group containing 1 to 6 carbon atoms.
  • Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl and tert-butyl), pentyl (n-pentyl, isopentyl and neopentyl) and hexyl.
  • the alkyl group may also be a part of another group, such as a C 1 -C 6 alkoxy group.
  • C 2 -C 6 alkylene refers to a divalent saturated aliphatic hydrocarbon group having 2 to 6 carbon atoms.
  • Alkylene groups include branched and straight chain hydrocarbyl groups.
  • (C 2 -C 6 )alkylene is meant to include ethylene, propylene, 2-methylpropylene, dimethylethylene, pentylene and the like.
  • C 2 -C 6 alkynylene group refers to a divalent aliphatic hydrocarbon group having 2 to 6 carbon atoms and containing one or more carbon-carbon triple bonds.
  • Alkynylene groups include branched and straight chain groups.
  • halogen refers to a halogen group: fluorine, chlorine, bromine or iodine.
  • the immune adjuvant in the present invention is an immunomodulator, which refers to a substance that has the function of enhancing and regulating immunity in terms of anti-infection, anti-virus, anti-tumor, anti-allergic reaction, anti-asthma, etc. It is mainly used clinically for adjuvant treatment of inflammation, autoimmune diseases, infectious diseases, cancer, and precancerous syndromes.
  • pharmaceutically acceptable refers to a certain carrier, vehicle, diluent, excipient, and/or the salt formed is usually chemically or physically compatible with other ingredients constituting a certain pharmaceutical dosage form, and physiologically compatible with the receptor.
  • salts and “pharmaceutically acceptable salts” refer to the above-mentioned compounds or their stereoisomers, or acid salts and/or basic salts formed with inorganic and/or organic acids and bases, and also include zwitterionic salts (inner salt), also include quaternary ammonium salts, such as alkyl ammonium salts. These salts can be directly obtained in the final isolation and purification of the compound. They can also be obtained by appropriately mixing the above-mentioned compound, or its stereoisomer, with a certain amount of acid or base (for example, equivalent).
  • salts may form a precipitate in the solution and be collected by filtration, or recovered after evaporation of the solvent, or prepared by freeze-drying after reacting in an aqueous medium.
  • the salt in the present invention can be the hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compounds.
  • one or more compounds of the present invention may be used in combination with each other.
  • the compound of the present invention can be used in combination with any other active agent to prepare drugs or pharmaceutical compositions for regulating cell function or treating diseases. If a group of compounds are used, these compounds can be administered to the subject simultaneously, separately or sequentially.
  • the compound provided by the present invention can effectively combine with STING, has a good STING protein agonistic function, shows a good inhibitory effect on a variety of tumors, and can also activate the immune memory mechanism of mice and inhibit tumor re-stimulation. Therefore, the compounds of the present invention can be used as STING agonists and used to treat various related diseases.
  • the compounds of the present invention have very good application prospects in the manufacture of a medicament for the treatment of diseases related to STING activity (especially medicaments for the treatment of inflammatory diseases, allergic diseases, autoimmune diseases, infectious diseases, cancer or precancerous syndromes) and in the manufacture of immune adjuvants, thus provide a new choice for clinical screening and/or manufacture of a medicament for treating diseases related to STING activity.
  • the structures of the compounds were determined by nuclear magnetic resonance (NMR) or (and) mass spectrometry (MS).
  • NMR shift ( ⁇ ) is given in unit of 10 -6 (ppm).
  • NMR was measured with (Bruker Avance III 400 and Bruker Avance 300) nuclear magnetic instrument, the solvent was deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated chloroform (CDCl 3 ), deuterated methanol (CD 3 OD), and the internal standard was tetramethylsilane (TMS).
  • DMSO-d 6 dimethyl sulfoxide
  • CDCl 3 deuterated chloroform
  • CD 3 OD deuterated methanol
  • TMS tetramethylsilane
  • the LC-MS was measured with Shimadzu LC-MS (Shimadzu LC-MS 2020 (ESI)).
  • the Shimadzu high pressure liquid chromatograph (Shimadzu LC-20A) was used for the HPLC measurement.
  • the hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1L.
  • the hydrogenation reaction is usually performed by vacuumizing, and filling with hydrogen, and the operations are repeated 3 times.
  • reaction is carried out under a nitrogen atmosphere.
  • the solution refers to an aqueous solution.
  • M is mole per liter.
  • Room temperature is the most suitable reaction temperature, which is 20°C-30°C.
  • PE refers to petroleum ether
  • EA refers to ethyl acetate
  • DCM refers to dichloromethane
  • MeOH refers to methanol
  • DMF refers to N,N-dimethylformamide
  • DMSO refers to dimethyl sulfoxide
  • DMAP 4-dimethylaminopyridine
  • DIPEA refers to diisopropylethylamine
  • Boc refers to tert-butyloxycarbonyl
  • TFA refers to trifluoroacetic acid
  • DBU refers to 1,8-diazabicycloundec-7-ene
  • HATU refers to 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate.
  • 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl chloride (4.46 g, 25.9 mmol) was dissolved in dry acetone (20 mL) and added dropwise to a solution of potassium thiocyanate (5.0 g, 51.5 mmol) in acetone (100 ml) and stirred at room temperature for 3 h.
  • the reaction system was filtered to remove inorganic salts.
  • Step 2 Synthesis of methyl (E)-3-amino-4-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-5-methoxybenzoate
  • Step 3 synthesis of methyl (E)-1-(4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamido)-7-methoxy-1H-methyl-benzo[d]imidazole-5-carboxylate
  • Step 5 Synthesis of methyl (E)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-methoxy-1-(4-((4-(methoxycarbonyl)-2-(3-morpholinopropoxy)-6-nitrophenyl)amino)but-2-en-1-yl)-1H-benzo[d]imidazole-5-carboxylate
  • Step 6 Synthesis of methyl (E)-1-(4-((2-amino-4-(methoxycarbonyl)-6-(3-morpholinopropoxy)phenyl)amino)but-2-ene-1-yl)-2-(4-ethyl)-2-methylthiazole-5-formylamino)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate
  • Step 7 synthesis of methyl (E)-2-(4-ethyl-2-methylthiazole-5-formylamino)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-5-(methoxycarbonyl)-7-(3-morpholinomethyl)-1H-benzo[d]imidazol-1-yl)but-2-ene-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate
  • Step 8 Synthesis of (E)-1-(4-(5-carboxy-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-(3-morpholinomethyl)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid
  • Step 9 synthesis of (E)-N-(5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-(3-morpholino)-1H--benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methylthiazole-5-formamide
  • Step 1 synthesis of methyl (E)-2-(4-ethyl-2-methylthiazole-5-formylamino)-1-(4-(2-(4-ethyl-2-methylthiazole-5-formylamino)-5-(methoxycarbonyl)-7-methyl ester-(3-morpholinomethyl)-1H-benzo[d]imidazol-1-yl)but -2-ene-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate
  • Step 2 synthesis of (E)-1-(4-(5-carboxy-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(3-morpholinomethyl)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid
  • Step 3 synthesis of (E)-N-(5-carbamoyl-1-(4-(5-carbamoyl-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(3-morpholinomethyl)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-2-yl)-4-ethyl-2-methylthiazole-5-formamide
  • Step 1 synthesis of methyl (E)-4-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-3-(3-morpholinopropoxy)-5-nitrobenzoate
  • Step 2 synthesis of methyl (E)-3-amino-4-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-5-(3-morpholinopropoxyl)benzoate
  • Step 3 synthesis of methyl (E)-1-(4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(3-morpholinomethyl)-1H-benzo[d]imidazole-5-carboxylate
  • Step 4 synthesis of methyl (E)-1-(4-aminobut-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(3-morpholinopropoxy)-1H-benzo[d]]imidazole-5-carboxylate
  • Step 6 synthesis of methyl (E)-1-(4-((2-amino-4-(methoxycarbonyl)-6-(methylthio)phenyl)amino)but-2-en-1-yl)-2-(4-ethyl-2)methyl ester-methylthiazole-5-formylamino)-7-(3-morpholinomethyl)-1H-benzo[d]imidazole -5-carboxylate
  • Step 7 Synthesis of methyl (E)-2-(4-ethyl-2-methylthiazole-5-formylamino)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-5-(methoxycarbonyl)-7-(methylthio)-1H-benzo[d] imidazol-1-yl)but-2-en-1-yl)-7-(3-morpholinomethyl)-1H-benzo[d]imidazole-5-carboxylate
  • Compound 5 was prepared by the same method as compound 4, except that the ring-closure materials used in step 3 and step 7 were exchanged, that is, 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate was used in step 3 for ring closure, and 4-ethyl-2-methylthiazole-5-carbonyl isothiocyanate was used in step 7 for ring closure.
  • the other steps were the same as the experiment method, and compound 5 (67 mg) was obtained.
  • Step 1 synthesis of methyl (E)-1-(4-((2-(3-(4-(tert-butoxycarbonyl)piperazin-1-yl)propoxy)-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-ketoen-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate
  • Step 2 synthesis of methyl (E)-1-(4-((2-amino-6-(3-(4-(tert-butoxycarbonyl)piperazin-1-yl)propoxy)-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-methoxy 1-H-benzo[d]imidazole-5-carboxylaye
  • Step 3 synthesis of methyl (E)-7-(3-(4-(tert-butoxycarbonyl)piperazin-1-yl)propoxy)-2-(4-ethyl-2-methylthiazole-5-formylamino)-1-(4-(2)-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-methoxy-5-(methoxycarbonyl)-1H-benzo[d]imidazole-1-yl)but-2-en-1-ylmethyl)-1H-benzo[d]imidazole-5-carboxylate
  • Step 4 synthesis of (E)-7-(3-(4-(tert-butoxycarbonyl)piperazin-1-yl)propoxy)-1-(4-(5-carboxy-2-(1-ethyl)-3-methyl-1H-pyrazole-5-formylamino)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-1H- -benzo[d]imidazole-5-carboxylic acid
  • Step 5 synthesis of tert-butyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole- 5-formylamino)-7-tert-butyl ester methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(4-ethyl -2-methylthiazole-5-formylamino)-1H-benzo[d]imidazol-7-yl)oxy)propyl) piperazine-1-carboxylate
  • Step 6 synthesis of (E)-N-(5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-(3-(piperazin-1-yl)propoxy)-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methylthiazole-5-formylamide
  • Step 7 synthesis of (E)-N-(5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-(3-(4-methylpiperazin-1-yl)propoxy)-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methylthiazole-5-formylamide
  • reaction solution was cooled to 0°C, then sodium cyanoborohydride (11 mg, 0.17 mmol) was added, reacted overnight at room temperature, and the reaction was quenched by adding water. After concentrating the reaction solution, it was purified by preparative HPLC to obtain compound 14 (23 mg, yield 43.7%).
  • Step 1 Synthesis of methyl (E)-methyl 2-(4-ethyl-2-methylthiazole-5-formylamino)-1-(4-(((2-methoxy-4-(methoxycarbonyl))-6-nitrophenyl)amino)but-2-en-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylate
  • Step 2 synthesis of methyl (E)-methyl 1-(4-(((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-carboxyamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylate
  • Step 3 synthesis of methyl (E)-methyl 2-(4-ethyl-2-methylthiazole-5-formylamino)-1-(4-(2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino)-7-methoxy-5-(methoxycarbonyl)-1H-benzo[d]imidazol-1-yl)but-2-ene-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylate
  • Step 4 synthesis of (E)-1-(4-(5-carboxy-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-carboxamino)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylic acid
  • Step 1 synthesis of methyl (E)-methyl 2-(4-ethyl-2-methylthiazole-5-formylamino)-1-(4-(2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino)-5-(methoxycarbonyl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)-but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate
  • Step 2 synthesis of (E)-1-(4-(5-carboxy-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-carboxamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-methylamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid
  • Step 3 synthesis of (E)-N-(5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino))-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methylthiazole-5-formylamide
  • Step 1 synthesis of methyl (E)-methyl 2-(4-ethyl-2-methylthiazole-5-formylamino)- 1-(4-(2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino)-5-(methoxycarbonyl)-7-(methylthio)-1H-benzo[d]imidazol-1-yl)but-2-ene-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole 5-carboxylate
  • Step 2 synthesis of (E)-1-(4-(5-carboxy-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino)-7-(methylthio)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylic acid
  • Step 3 synthesis of (E)-N-(5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino))-7-(methylthio)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methylthiazole-5-formylamide
  • Step 1 synthesis of methyl (E)-methyl 2-(4-ethyl-2-methylthiazole-5-formylamino)-1-(4-(2-(4-ethyl-2-methylthiazole-5-formylamino)-5-(methoxycarbonyl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-(methylthio)-1H-benzo[d]imidazole-5-carboxylate
  • Step 2 synthesis of (E)-1-(4-(5-carboxy-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(methylthio)-1H-benzo[d]imidazole-5-carboxylic acid
  • Step 3 synthesis of (E)-1-(4-(5-carboxy-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(methylthio)-1H-benzo[d]imidazole-5-formylamide
  • Step 1 synthesis of methyl (E)-methyl 1-(4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-carboxamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylate
  • Step 2 synthesis of methyl (E)-methyl 1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylate
  • Step 3 synthesis of methyl (E)-methyl 2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino)-1-(4-((4-(methoxycarbonyl)-2-(methylthio)-6-nitrophenyl)amino)but-2-en-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylate
  • Step 4 synthesis of methyl (E)-1-(4-((2-amino-4-(methoxycarbonyl)-6-(methylthio)phenyl)amino)but-2-en-1-yl)-2-(4-ethyl-2)methyl ester-methylthiazole-5-formylamino)-7-(3-morpholinomethyl)-1H-benzo[d]imidazole -5-carboxylate
  • Step 6 synthesis of (E)-1-(4-(5-carboxy-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-(methylthio)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-carboxamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylic acid
  • Step 7 synthesis of (E)-1-(4-(5-(5-carbamoyl-2-)(1-styrene-3-methyl-1H-pyrazole-5-formylamino)-7-(methylthio)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-styrene-4-fluoro-3-methyl-1H-pyrazole-5-anthralamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-formylamide
  • Step 2 synthesis of (E)-1-(4-(5-carboxy-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-carboxamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-(methylthio)-1H-benzo[d]imidazole-5-carboxylic acid
  • Step 3 synthesis of (E)-N-(5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino))-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-(methylthio)-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methylthiazole-5-formylamide
  • Step 1 synthesis of methyl (E)-methyl 2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-carboxamino)-1-(4-(2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino)-5-(methoxycarbonyl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)-2-en-1-yl)-7-(methylthio))-1H-benzo[d]imidazole-5-carboxylate
  • Step 2 synthesis of (E)-1-(4-(5-carboxy-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-carboxamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino)-7-(methylthio)-1H-benzo[d]imidazole-5-formamide
  • Step 3 synthesis of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-carboxamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-formylamino)-7-(methylthio)-1H-benzo[d]imidazole-5-carboxamide
  • Step 1 synthesis of methyl (E)-methyl 1-(4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylate
  • Step 2 synthesis of methyl (E)-methyl 1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylate
  • Step 3 synthesis of methyl (E)-methyl 2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-1-(4-((4-(methoxycarbonyl)-2-(methylthio)-6-nitrophenyl)amino)but-2-en-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylate
  • Step 4 synthesis of methyl (E)-methyl 1-(4-(((2-amino-4-(methoxycarbonyl)-6-(methylthio)phenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxylate
  • Step 5 synthesis of methyl (E)-methyl 2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-5-(methoxycarbonyl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)butan-2-en-1-yl)-7-(methylthio)-1H-benzo[d]imidazole-5-carboxylate
  • Step 6 synthesis of (E)-1-(4-(5-carboxy-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamino)-7-(methylthio)-1H-benzo[d]imidazole-5-carboxylic acid
  • Step 7 synthesis of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-y1)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamino)-7-(methylthio)-1H-benzo[d]imidazole-5-formylamide
  • Step 2 synthesis of tert-butyl (E)-(4-((4-carbamoyl-5-fluoro-2-methoxyphenyl)amino)but-2-en-1-yl)carbamate
  • Step 3 synthesis of tert-butyl (E)-(4-((4-carbamoyl-5-fluoro-2-methoxy-6-nitrophenyl)amino)but-2-en-1-yl)carbamate
  • Step 4 synthesis of tert-butyl (E)-(4-((4-carbamoyl-5-fluoro-2-methoxy-6-aminophenyl)amino)but-2-en-1-yl)carbamate
  • Step 5 synthesis of tert-butyl (E)-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-4-fluoro-7-methoxy-1H-benzo[d]imidazol-1-yl)-2-en-1-yl)carbamate
  • Step 6 synthesis of methyl (E)-1-(4-aminobut-2-en-1-yl)-2-(4-ethyl-2-methylthiazole-5-formylamino)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate
  • Step 7 synthesis of (E)-1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-4-fluoro-7-methoxy-1H-benzo[d]imidazole-5-carboxamide
  • Step 8 synthesis of (E)-1-(4-(((2-amino-4-carbamoyl-6-(3-morpholinopropoxy) phenyl)amino)but-2-ene-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)-4-fluoro-7-methoxy-1H-benzo[d]imidazole-5-formylamide
  • Step 9 synthesis of (E)-N-(5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-formylamino)yl)-4-fluoro-7-methoxy-1H)-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methylthiazole-5-formylamide
  • the protein thermal transfer test was used to determine the binding affinity of the compound to Sting protein.
  • the protein dissolution curve was measured on a qPCR instrument, the Tm value was fitted with Protein Thermal Shift Software 1.3 software, and the Tm difference of the protein was calculated when different concentrations of compound and no compound were added.
  • the dissociation constant Kd was fitted and obtained according to the change of ⁇ Tm with the concentration of the compound.
  • the compound 3 reported in the article ( Nature. 2018 Dec;564(7736):439-443 ) was used as a positive reference compound.
  • the test results are shown in Table 5 below.
  • the Kd value of each compound is classified according to the following instructions:
  • the function of sting agonist was evaluated by detecting the changes of IFN- ⁇ and CXCL10 (IP10) cytokines produced by human peripheral blood mononuclear cell line THP1 cells (Shanghai Cell Bank) stimulated by the compounds.
  • the ELISA plate was coated according to the IFN- ⁇ (R&D, #DY814-05) and IP10 (BD, #550926) ELISA test kit instructions. The compound was dissolved into DMSO stock solution and diluted with culture medium to a 2X working concentration, added to a 96-well plate, 100 ⁇ L per well.
  • the THP1 cells in the logarithmic growth phase were taken to count, and were diluted to a concentration of 2* 10 6 /mL, added to the above-mentioned 96-well plate containing the compounds, with 100 ⁇ L per well, mixed well, and incubated in a 37°C, 5% CO 2 incubator for 18 hours.
  • the above cell culture supernatant was taken, 100 ⁇ L per well, and tested according to IFN- ⁇ and IP10 ELISA test kits respectively.
  • the OD450 value was read, and converted into IFN- ⁇ and IP10 concentration according to the standard curve, and fitted with GraphPad 5.0 to calculate the EC 50 value of the dose-efficiency curve.
  • EC 50 is the half-maximal effect concentration (concentration for 50% of maximal effect, EC 50 ), which refers to the concentration of the drug that can cause 50% of the individual to be effective.
  • the above experimental data show that the compounds of the present invention have good stimulating activity of IFN- ⁇ and CXCL10 (IP10) cytokines produced by THP1 cells, and have a good STING protein agonistic function. Especially for compounds 2, 4, 5, 15, 19, 23, 35, 40, 49, 50, 51, 52, 53, etc., compared with the reference compound, the compounds of the present invention have comparable or even higher activity of stimulating the immune cells to produce cytokines.
  • IP10 CXCL10
  • mice were inoculated with 5 ⁇ 10 5 CT26 cells subcutaneously on the back. After inoculation, tumor-bearing mice with tumor volume in the range of 200-300 mm 3 were selected for the experiment. According to the random method of tumor size and segment, they were divided into 21 groups with 3 mice in each group. Different doses of drugs (compounds of the present invention) were given via tail vein injection, and the control group was given a control vehicle (5% DMSO, 40% PEG400 formulated in physiological saline). Three hours after the administration, blood was taken from the mouse orbital vein, and EDTA•2K was used as an anticoagulant. The collected mouse whole blood was centrifuged at 8000g for 5 minutes and then the upper plasma was taken for testing.
  • drugs compounds of the present invention
  • mouse tumor tissues were taken, weighed and homogenized by adding 9 times volume of PBS. After centrifuging the homogenate at 10,000 rpm for 15 minutes, the supernatant was collected for testing. The detection was performed according to the IFN- ⁇ detection kit (R&D, #DY814-05), the OD450 value was read, which was converted to the IFN- ⁇ concentration according to the standard curve.
  • mice can induce the release of IFN- ⁇ downstream of the STING pathway in blood and tumors.
  • the intratumor IFN- ⁇ level was more than 3 times that of the plasma IFN- ⁇ level, indicating that cytokines were relatively enriched in tumor tissues, and blood cytokines were low, suggesting a higher treatment window.
  • mice After Balb/c mice rested for one week, CT26 cells were inoculated subcutaneously on the back, and each mouse was inoculated with 100 ⁇ L cell suspension containing 5 ⁇ 10 5 cells. When the average tumor volume grew to about 120 mm 3 , they were randomly divided into groups according to the size of the tumor, with 5 in each group. The mice in each group were administered intravenously through the tail vein on Day 1, 4, and 8 after grouping, and the control group was given a control vehicle (5% DMSO, 40% PEG400 formulated in physiological saline).
  • 5% DMSO, 40% PEG400 formulated in physiological saline 5% DMSO, 40% PEG400 formulated in physiological saline.
  • the length and short diameter of the tumor were measured and recorded with a vernier caliper, the tumor volume was calculated, and the tumor growth curve was drawn according to the tumor volume.
  • Kunming rats were inoculated with H22 cells subcutaneously on their backs after a week of resting. Each mouse was inoculated with 100 ⁇ L of cell suspension containing 5 ⁇ 10 6 cells. When the average tumor volume grew to about 200 mm 3 , they were randomly grouped according to the tumor size, with 7 in each group. On the Day 1, 4 and 8 after grouping, the mice in the high, medium and low dose groups were administered intravenously via the tail vein (the compounds of the present invention), or on the Day 1, 8, and 15 the mice in the high dose group were administered intravenously via the tail vein. The control group was given a control vehicle (5% DMSO, 40% PEG400 formulated in physiological saline).
  • the length and short diameter of the tumor were measured and recorded with a vernier caliper, the tumor volume was calculated, and the tumor growth curve was drawn according to the tumor volume.
  • the compounds of the present invention After 30 days of administration, the compounds of the present invention all effectively inhibited tumor growth, and exhibited a dose-dependent tumor growth inhibitory effect.
  • the experimental results are shown in Figure 2 .
  • the CT26 tumors in the control group can grow after inoculation, but the tumors in the mice whose tumors disappeared completely after treatment with the compounds of the present invention cannot grow again after inoculation, indicating that these compounds activated the immune memory mechanism of the mice, and render mice had immunity to tumor cells that were re-inoculated.
  • the present invention discloses a compound which is capable of effectively binding to STING, has a good STING protein agonistic function, has a good inhibitory effect on a variety of tumors, and can also activate immune memory mechanism in mice, to inhibit tumor restimulation. Therefore, the compounds of the present invention can be used as STING agonists and used to treat various related diseases.
  • the compounds of the present invention have very good application prospects in the manufacture of a medicament for the treatment of diseases related to STING activity (especially a medicament for the treatment of inflammatory diseases, allergic diseases, autoimmune diseases, infectious diseases, cancer or precancerous syndromes) and in the manufacture of immune adjuvants, thus provides a new choice for clinical screening and/or manufacture of a medicament for diseases related to STING activity.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Immunology (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Oncology (AREA)
  • Pain & Pain Management (AREA)
  • Rheumatology (AREA)
  • Communicable Diseases (AREA)
  • Pulmonology (AREA)
  • Transplantation (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Plural Heterocyclic Compounds (AREA)
  • Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Claims (7)

  1. Verbindung oder das Stereoisomer oder das pharmazeutisch verträgliche Salz davon, wobei die Verbindung durch die Formel II dargestellt ist, wobei X aus -O- und -S- ausgewählt ist und X' -O- ist;
    Ring A und Ring A' jeweils unabhängig voneinander ausgewählt sind aus optional substituiert durch 0-2 Rc; und wobei, wenn X und X' beide -O- sind, mindestens einer von Ring A und Ring A' ausgewählt ist aus optional substituiert durch 0-2 Rc;
    R5 ausgewählt ist aus C1-C3-Alkyl;
    R5' ausgewählt ist aus
    Re' ausgewählt ist aus C1-C3-Alkyl, 3-6-gliedrigem Cycloalkyl, -C(O)Rf;
    Rf ausgewählt ist aus -ORa', C1-C3-Alkyl, 3- bis 4-gliedrigem Cycloalkyl, und Ra' ausgewählt ist aus C1-C3-Alkyl; und
    wobei
    das optional substituiert durch 0 bis 2 Rc, oder ist; und
    das optional substituiert durch 0-2 Rc, ist.
  2. Verbindung oder das Stereoisomer oder das pharmazeutisch verträgliche Salz davon, wobei die Verbindung Folgendes ist:
  3. Verbindung oder das Stereoisomer oder das pharmazeutisch verträgliche Salz davon, wobei die Verbindung durch die Formel III dargestellt ist: wobei R1, R3, R1' und R3' Wasserstoff sind;
    wobei Ring A und Ring A' jeweils unabhängig voneinander ausgewählt sind aus und mindestens einer von Ring A und Ring A' ist;
    R5' ausgewählt ist aus C1-C3-Alkyl, optional substituiert durch 0-2 Rd;
    Rd unabhängig ausgewählt ist aus
  4. Verbindung oder das Stereoisomer oder das pharmazeutisch verträgliche Salz davon, wobei die Verbindung Folgendes ist:
  5. Verbindung oder das Stereoisomer oder das pharmazeutisch verträgliche Salz davon nach einem der Ansprüche 1 bis 4 zur Verwendung bei der Behandlung einer Krankheit, die mit der STING-Aktivität in Zusammenhang steht;
    wobei die mit der STING-Aktivität in Zusammenhang stehende Krankheit eine der Krankheiten ist, die mit Entzündungskrankheiten, Autoimmunerkrankungen, Infektionskrankheiten, Krebs und präkanzerösen Syndromen oder einer Kombination davon in Zusammenhang stehen.
  6. Verbindung oder das Stereoisomer oder das pharmazeutisch verträgliche Salz davon nach einem der Ansprüche 1 bis 4 zur Verwendung als Immunadjuvans.
  7. Arzneimittel, das eine Formulierung ist, die mit der Verbindung oder dem Stereoisomer oder dem pharmazeutisch verträglichen Salz davon nach einem der Ansprüche 1 bis 4 als Wirkstoff sowie pharmazeutisch verträglichen Hilfsstoffen hergestellt ist.
EP20748337.1A 2019-01-31 2020-01-21 Immunmodulator Active EP3919488B1 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
CN201910083149 2019-01-31
CN201910819220 2019-09-04
PCT/CN2020/073405 WO2020156363A1 (zh) 2019-01-31 2020-01-21 一种免疫调节剂

Publications (4)

Publication Number Publication Date
EP3919488A1 EP3919488A1 (de) 2021-12-08
EP3919488A4 EP3919488A4 (de) 2022-04-06
EP3919488C0 EP3919488C0 (de) 2025-08-27
EP3919488B1 true EP3919488B1 (de) 2025-08-27

Family

ID=71840242

Family Applications (1)

Application Number Title Priority Date Filing Date
EP20748337.1A Active EP3919488B1 (de) 2019-01-31 2020-01-21 Immunmodulator

Country Status (9)

Country Link
US (1) US20220119379A1 (de)
EP (1) EP3919488B1 (de)
JP (1) JP7655854B2 (de)
KR (1) KR102702618B1 (de)
CN (2) CN113563313B (de)
AU (1) AU2020214477B2 (de)
CA (1) CA3128069C (de)
ES (1) ES3044732T3 (de)
WO (1) WO2020156363A1 (de)

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2020324388A1 (en) * 2019-08-02 2022-02-24 Mersana Therapeutics, Inc. Bis-[N-((5-carbamoyl)-1H-benzo[d]imidazol-2-yl)-pyrazol-5-carboxamide] derivatives and related compounds as STING (Stimulator of Interferon Genes) agonists for the treatment of cancer
EP4126065A1 (de) 2020-04-02 2023-02-08 Mersana Therapeutics, Inc. Antikörper-wirkstoff-konjugate mit sting-agonisten
KR20220167275A (ko) 2020-04-10 2022-12-20 오노 야꾸힝 고교 가부시키가이샤 암 치료 방법
BR112023014128A2 (pt) * 2021-01-15 2023-10-31 Seagen Inc Conjugados de anticorpo-fármaco imunomodulatórios
US12065427B2 (en) * 2021-04-29 2024-08-20 Boehringer Ingelheim International Gmbh Heterocyclic compounds capable of activating STING
WO2022246597A1 (en) * 2021-05-24 2022-12-01 Forever Millets Limited Imidazopyridine derivatives as sting agonists
CN113429387B (zh) * 2021-07-27 2022-10-28 中国药科大学 一种苯并[b]硒吩类STING调控剂、其制备方法及用途
CN115724838A (zh) * 2021-08-26 2023-03-03 成都先导药物开发股份有限公司 一种适合作为抗体偶联药物效应分子的sting激动剂
JP2025502881A (ja) * 2022-01-11 2025-01-28 ビシケム カンパニー リミテッド Ｓｔｉｎｇ作用剤としての縮合ヘテロアリールヒドロキサメート
AU2024315750A1 (en) 2023-07-31 2026-03-05 Sutro Biopharma, Inc. Sting agonist compounds and conjugates
US20260053942A1 (en) 2024-05-30 2026-02-26 Astellas Pharama Inc. Sting agonist immunostimulatory antibody drug conjugates

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
PL3440076T3 (pl) 2016-04-07 2022-09-05 Glaxosmithkline Intellectual Property Development Limited Heterocykliczne amidy przydatne jako modulatory białek
WO2019069269A1 (en) * 2017-10-05 2019-04-11 Glaxosmithkline Intellectual Property Development Limited INTERFERON GENE STIMULATOR MODULATORS USEFUL IN THE TREATMENT OF HIV
CA3077337A1 (en) * 2017-10-05 2019-04-11 Glaxosmithkline Intellectual Property Development Limited Modulators of stimulator of interferon genes (sting)
CN110016025B (zh) * 2018-01-08 2021-08-06 成都先导药物开发股份有限公司 一种免疫调节剂
AU2020324388A1 (en) * 2019-08-02 2022-02-24 Mersana Therapeutics, Inc. Bis-[N-((5-carbamoyl)-1H-benzo[d]imidazol-2-yl)-pyrazol-5-carboxamide] derivatives and related compounds as STING (Stimulator of Interferon Genes) agonists for the treatment of cancer
CN115724838A (zh) * 2021-08-26 2023-03-03 成都先导药物开发股份有限公司 一种适合作为抗体偶联药物效应分子的sting激动剂

Also Published As

Publication number Publication date
EP3919488C0 (de) 2025-08-27
WO2020156363A1 (zh) 2020-08-06
CN113563313A (zh) 2021-10-29
CA3128069C (en) 2023-10-10
JP7655854B2 (ja) 2025-04-02
KR102702618B1 (ko) 2024-09-06
KR20210120070A (ko) 2021-10-06
EP3919488A1 (de) 2021-12-08
CN111499617A (zh) 2020-08-07
ES3044732T3 (en) 2025-11-27
CN113563313B (zh) 2022-11-04
EP3919488A4 (de) 2022-04-06
JP2022518853A (ja) 2022-03-16
US20220119379A1 (en) 2022-04-21
CA3128069A1 (en) 2020-08-06
CN111499617B (zh) 2021-08-06
AU2020214477A1 (en) 2021-09-02
AU2020214477B2 (en) 2022-10-27

Similar Documents

Publication Publication Date Title
EP3919488B1 (de) Immunmodulator
EP3743420B1 (de) Heterocyclylaminosubstituierte triazole als modulatoren der rho-assoziierten proteinkinase
AU2019342102B2 (en) Compounds for treating certain leukemias
ES2620316T3 (es) Cicloalquenopirazoles sustituidos como inhibidores de BUB1 para el tratamiento del cáncer
WO2023138524A1 (zh) Kras g12d降解剂及其在医药上的应用
KR20240136407A (ko) 벤조피리미딘 화합물 및 이의 용도
CN114409653A (zh) 一种桥环并嘧啶并环类化合物及其用途
EP3068785A1 (de) Substituierte 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-derivative als casein-kinase-1-d/e-inhibitoren
CN105452242A (zh) 杂芳基取代的吡唑
CN111606861A (zh) 用于治疗病毒感染和另外的疾病的酰氨基嘧啶衍生物
JP2003514804A (ja) ファルネシルトランスフェラーゼ阻害剤としてのトリアゾール類
EP2315521A1 (de) Diaminopyridin-, pyrimidin- und pyridazinmodulatoren des histamin-h<sb>4</sb>-rezeptors
EP3873474A1 (de) Heterocyclische verbindungen als bet-hemmer
CN105473570A (zh) 二氨基杂芳基取代的吡唑
AU2020301451B2 (en) Tricyclic compound, preparation method therefor and use thereof
KR20200090636A (ko) 피롤로피리미딘 유도체 및 이를 유효성분으로 함유하는 단백질 키나아제 관련 질환의 예방 또는 치료용 약학적 조성물
JP2025514382A (ja) Ｒｏｃｋ２の阻害剤
CN114599655B (zh) 咪唑烷酮类化合物及其制备方法与应用
EP4242213A1 (de) Borsäureverbindung
KR102628246B1 (ko) 선택성 a2a 수용체 대항제
CN117447460A (zh) 作为ripk1抑制剂的杂环化合物
EP2578588A1 (de) Neue 1,4-diazepam-pde-5-hemmer-derivate
HK40036815A (en) Acylaminopyrimidine derivatives for the treatment of viral infections and further diseases
HK40061722B (zh) 杂环酰胺类化合物及其制备方法和应用
HK1157569B (en) Diamino-pyridine, pyrimidine, and pyridazine modulators of the histamine h 4 receptor

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20210816

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

REG Reference to a national code

Ref country code: DE

Ref legal event code: R079

Free format text: PREVIOUS MAIN CLASS: C07D0405140000

Ipc: C07D0417140000

Ref document number: 602020057426

Country of ref document: DE

A4 Supplementary search report drawn up and despatched

Effective date: 20220310

RIC1 Information provided on ipc code assigned before grant

Ipc: A61P 35/00 20060101ALI20220303BHEP

Ipc: A61P 31/00 20060101ALI20220303BHEP

Ipc: A61P 29/00 20060101ALI20220303BHEP

Ipc: A61P 37/00 20060101ALI20220303BHEP

Ipc: A61K 31/427 20060101ALI20220303BHEP

Ipc: C07D 491/107 20060101ALI20220303BHEP

Ipc: C07D 491/048 20060101ALI20220303BHEP

Ipc: C07D 405/14 20060101ALI20220303BHEP

Ipc: C07D 403/14 20060101ALI20220303BHEP

Ipc: C07D 417/14 20060101AFI20220303BHEP

DAV Request for validation of the european patent (deleted)
DAX Request for extension of the european patent (deleted)
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

17Q First examination report despatched

Effective date: 20230523

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: GRANT OF PATENT IS INTENDED

INTG Intention to grant announced

Effective date: 20250327

GRAS Grant fee paid

Free format text: ORIGINAL CODE: EPIDOSNIGR3

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE PATENT HAS BEEN GRANTED

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

REG Reference to a national code

Ref country code: CH

Ref legal event code: EP

REG Reference to a national code

Ref country code: IE

Ref legal event code: FG4D

U01 Request for unitary effect filed

Effective date: 20250902

U07 Unitary effect registered

Designated state(s): AT BE BG DE DK EE FI FR IT LT LU LV MT NL PT RO SE SI

Effective date: 20250908

REG Reference to a national code

Ref country code: ES

Ref legal event code: FG2A

Ref document number: 3044732

Country of ref document: ES

Kind code of ref document: T3

Effective date: 20251127

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IS

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20251227

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: NO

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20251127

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: HR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20250827

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: GR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20251128

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: PL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20250827

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: RS

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20251127

REG Reference to a national code

Ref country code: CH

Ref legal event code: U11

Free format text: ST27 STATUS EVENT CODE: U-0-0-U10-U11 (AS PROVIDED BY THE NATIONAL OFFICE)

Effective date: 20260201

U20 Renewal fee for the european patent with unitary effect paid

Year of fee payment: 7

Effective date: 20260115